168 resultados para Tetrazolium Salts
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The inhibitory effect of 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) and 2,4,6-tri(2-pyridyl)-s-triazine (TPT) molecules on the corrosion of mild steel in 1 mol/L HCl and microcosmic inhibitory mechanism were investigated by X-ray photoelectron spectroscopy and ellipsometry. XPS results showed that C Is and N Is peaks of TTC, C Is and N Is peaks of TPT and their integral areas were obtained, which suggested the layer of the inhibitors (TTC or TPT) should have effectively protected the mild steel surface from the corrosion; and the depression from the inhibitors for the corrosion of mild steel surface was studied using ellipsometry combined with potentiodynamic polarization and the phasic difference was gained, which displayed the inhibitory coverage of the inhibitors formed.
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Electrochemical measurement, quantum chemical method, and scanning electron microscopy (SEM) were performed to investigate the inhibitive effect of 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) and 2,4,6-tri(2-pyridyl)-s-triazine(TPT) on the corrosion of mild steel in 1mol.L-1 HCl at room temperature. Impedance spectroscopy measurement showed that the polarization resistance increased and that double layer capacitance decreased with the increase in the inhibitive concentration, and the results of potentiodynamic polarization showed that the inhibitors suppressed both cathodic and anodic processes of steel corrosion without change in the mechanism. Higher the orbital density distribution strength of the lowest unoccupied molecular orbital, higher is the molecule dipole, and lower energy gap between the energy of the highest occupied molecular orbital and the energy of the lowest unoccupied molecular orbital resulted in higher inhibitory efficiency. The results of SEM analysis showed that the metal was protected from aggressive corrosion by the addition of TTC and TPT.
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通过优化熔融条件和玻璃组份,成功开发出一种新的Er^3+/Yb^3+共掺磷酸盐玻璃,其在沸水和熔盐中均表现出很好的化学稳定性。通过分析室温下Er^3+/Yb^3+共掺磷酸盐玻璃的吸收光谱,计算得到了Er^3+离子在波长1533nm处的峰值发射截面和杜得-奥菲而特强度参数;其中Er^3+离子在波长1533nm处的峰值发射截面为0.72×10^20cm^2,大于Schott的IOG1玻璃中Er^3+离子的峰值发射截面0.67×10^-20cm^2。通过改变离子交换的条件,获得了1.55μm单模光波导的制作条件
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利用发根农杆菌(Agrobacterium rhizogenes)1601,1000,1500,15834,A4,均成功地转化了中药青蒿(Artemisia annua L.)并且建立了pRi1601,pRi15834,pRiA4诱导的发根培养。pRi1601,pRi15834的发根诱导率比其它质粒高。太老或太幼的叶片不利子发根的诱导;发根主要从叶脉的伤口处萌发;带顶芽或带侧芽的叶片容易诱导根,但不一定是发根。光照有利于发根的诱导和发根的生长。以每个发根的“绝对生长速率”(Gtowth Ratio,GR)和绝对“侧根”数量(Number of Side Roots,NSR),通过大量的发根系的筛选,建立了8个发根系,1601-L-1, 1601-L-2, 1601-L-3, 1601-L-4, 15834-L-1, 1601-P-I, 16 01-P-2,15834-L-2。Southern分子检测表明,160l-1-1,1801-L-2, 1601-L-3,1601-L-4,1601-P-1,1601-P-2均为转化子。8个建立的发根系之间无论生长或者QHS的合成存在明显的差异。比较光/暗(16/8hrs),25℃条件下培养的16 01-L-1,1601-L-2,1601-L-3,1601-L-4,1601-P-l,和1601-P-2,其中16 01-L-3的生长最快,160l-L-1的生长最慢;但是,1601-L-1的QHS的含量最高(可达1. 048%),1601-1-3的QHS的含量最低。160Z-L-3,15834 -L-1和2583:1-L-2的生长速率相差不大。用盛有l000mLMS液体培养基的3000mL的锥形瓶扩大培养1601-L -3,15834-L-1和15834-L-2,转速为ll0rlpm,培养过程中发根容易形成发根球(Hairy Root Balis,HRB),HRB的形成严重影响发根的生长和QHs的合成,HpLC分析表明扩大培养发根中QHS的含量比较低。 改变MS基本培养基中的无机离子的浓度,研究不同无机离子对发根生长和QHS的合成的影响。 l、KN03为18.79×10-3M时有利于1601- L-1生长,为14. 84×10-3M时有利于QHS的合成。NH-4N0-3浓度在10.93-12. 49×10—3M范围内有利于1601-L-1生长,在0-20.62×10-3M范围内对QHS的合成影响不大,大于20. 62×lO-3M不利QHS的合成。培养基中NH-4+/N0-3-比值为0. 37-0. 4-0.52:1时有利于发根的生长,比值为0.52 - 0.58:1时有利于QHS的合成。 2、H-2P0-4-浓度为2.498×10-3M时有利于发根的生长在0-2. 498×l0-3M范围内,随着浓度的提高,促进发根的生长。培养基中的H2P4 -的浓度在0-1.249×lO-3M的范围内,随着浓度的提高,促进QHS的合成,为1.249×10-3M时QHS的含量最高。 3、培养基中最适16 01-L-1生长的Ca-2+浓度为0.198- 0.766×10-3M,大于或小于该浓度范围,显著地抑制发根的生长。但是,在0-3.695×10-3M范围内,随着培养基中Ca-2+浓度提高,促进QHS的合成,最适Ca-2+浓度为3.695×l0-3M。 4、培养基中不加Mg-2+时,完全抑制发根生长,在0. 142×10-3M-7.506×l0-3M浓度范围内,对发根生长影响没有明显的差别。但是,HPLC和UV分析发根中QHS含量,培养基中不加Mg-2+时,发根中QHS含量最高。 5、培养基中的Fe-2+浓度在0. 25 -1.0×10-3M范围内,同时有利于16 01- L-1的生长和QHS的形成。 6、培养基中最适合予16 01- L-3生长的KI浓度为2.5ppm,大于或小予该浓度均显著地抑制发根的生长,培养基中加入KI明显地降低发根中的QHS的含量。 7、H2BO3对l601-L-l生长影响不大,HPLC分析QHS的含量,培养基中的H3BO3浓度为100ppm和400ppm,QHS的含量分别为1.69mg/g和1.80mg/g(DW)。 8、Cu-2+对1601-L-3的生长影响显著,最适合1601-L-3生长的Cu-2+浓度为1.00ppm,在0 -1.00ppm的浓度范围内,随着培养基中的Cu+浓度的提高,发根的生物量不断增加。培养基中QHS合成的最适Cu2+浓度为0.05ppm,大于或小于该浓度均显著地抑制发根中QHS的合成。 比较光培养和暗培养对发根生长的影响,结果表明光照明显地促进1601-L-l的生长,暗培养明显不利于发根的生长。最适合于发根生长的温度为25℃,大于35℃显著地抑制发根的生长,影响发根的根尖细胞的正常分裂。 改变培养基中的蔗糖浓度和在发根培养的不同时期给培养基中添加蔗糖,试验结果表明蔗糖作为碳源对1601-L-3和1601-L-1的生长具有显著的影响。 (1)培养基中缺少蔗糖显著地抑制发根的生长。 (2)发根培养的前5天时间内,蔗糖浓度为30- 60glL昀培养基最有利于发根的生长,50glL的培养基中的发根生长最快,培养基中的蔗糖浓度大于60g/L小于30g/L时,发根的生物量增加较少。 (3)发根培养至第15天时,蔗糖浓度为60g/L的培养基最有利予发根的生物量的增加。发根培养至30天时,蔗糖浓度为60-90g/L的培养基,发根的生物量的增加相差不大,但是为蔗糖浓度为30-40g/L的培养基中的发根生物量一倍。 (4)发根培养过程中,分别于第5和15天给蔗糖浓度为30g/L的培养基中添加一次或二次蔗糖,使培养基中的蔗糖终浓度相当于60g/L或90g/L,培养至30天时,添加蔗糖的培养基中的发根的干重生物量相当于不添加蔗糖培养基中的发根生物量一倍,相当于初始蔗糖浓度为60g/L和90g/L培养基中发根的生物量。 (5)随着培养基中蔗糖浓度的提高,发根干重/鲜重比显著增加。培养基中的蔗糖的消耗量与发根生物量的增加呈正相关,蔗糖消耗越多,发根生物量的增加越大。 比较pH值对发根生长和QHS合成的影响表明,灭菌前pH值在5.O-6.5范围内的培养基适合予1601-L-1的生长,小于5.O不利于发根的生长,pH5.8有利于1601-1-1生长和QHS的生物合成。发根收获时培养基中的pH值一般为4.5-5.2. pH7.O抑制发根的生长,pHl0.O对发根具有强烈的致死作用。发根在培养过程中,对培养基中的pH值具有显著的调节作用,发根能在很短的时间内(24- 48hrs)使pl:l值为5.8、6.4、7.0培养基降低到pH4. 5-5.2,pH为5.8的培养基有利于QHS合成。 比较不同基本培养基对发根生长和QHS合成的影响,试验结果表明N6、DCR、Litvay培养基有利于1601-L-1的生长,WS、White、B5培养基不利于发根的生长。DCR培养基中的QHS含量最高。 根据三水平试验选用三水平正交表来安排试验的原则,选用三水平正交表L7(3-),研究多因子效应对发根生长和QHS合成的影响,试验结果表明,Mg2+,Fe2+,Mn-2+,NH4NO3,KN03 ,KI,Ca-2+为发根生长的主要因子,NH4N03,KNOs,Mg2+,Ca2+,肌醇为QHS合成的主要因子。 通过TLC分析发根中QHS和其它化学成分,同时比较发根和无菌苗及野生植株的化学成分,发根和无菌苗均能合成包括QHS在内的野生青蒿叶片中的大部分非挥发性的化台 物。 研究青蒿植株在发育过程中QHS的含量的变化以及发根、无菌苗和野生青蒿中QHS的合成,HP分析结果表明,l、不同的单株青蒿之间的QHS量相差很大。2、同一植株幼 叶的QHS含量比老叶的QHS含量高。3、不同单株青蒿之间达到最高QHS含量的时间不一样,开花期或开花之前。4、无菌苗(带根)或者不带根丛生芽均能合成QHS,但是带根的无菌蕾的QHS量比丛生芽中的QIS的含量高。5、不同发根农杆菌转化的发根系1601-L-1和15834-L-1都能合成QHS。
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一.应用高通量技术筛选松树抗微生物分子的研究 我们提出一种对纯化组分的抗菌活力进行高通量技术筛选的比色方法。该方法利用一种新型四氮唑盐MTS(methyl tetrazolium salt)在电子偶联剂PMS( phenazine methosulfate)的辅助下对供试样品进行比色分析,其原理是MTS能穿透进完整活性细胞,在接受电子偶联剂PMS提供的电子后,被其中细胞器膜或质膜上的脱氢酶还原成一种水溶性的显色甲替(formazan)化合物。甲替在490nm波长的吸收值能直接用96孔酶标板读取,无须通过额外的溶解步骤。甲替的产生量(吸收值)和基质中的活细胞数成正比。因此,抗菌成分作用于供试细菌后,其抗菌活力大小可通过对照孔(未加抗菌成分)和试验孔吸收值的差异反映出来。借助该技术和对影响MTS转化的各种因素标准化,定量研究6种细菌菌株包括革兰氏阴性菌和革兰氏阳性菌的生长。通过反相C18柱浓缩、凝胶过滤层析和亲和层等技术,从受马尾松毛虫危害的马尾松针内分离和纯化出几种增强的抗微生物活力,证实了。该方法对临床供试菌株的可行性和应用潜力;同时,对植物宿主受害后的诱导生理和功能意义亦进行了探讨。 二.向日葵种质资源相关基础研究中文摘要 1) 维生素E的天然产物有八种类型,分别为α、β、γ、δ一生育酚( tocopherol)和α、β、γ、δ一生育三烯酚( tocotrienol),对植物、动物和人类都具有十分重要的生理作用。绿色植物是人类和动物VE的基本来源。本文对有关植物中VE生物合成途径和相关酶基因克隆研究现状,以及VE在植物体内的作用和功能研究进展进行了综述,以期为VE的机理探寻和功能开发提供进一步的思路。 2) 以20份向日葵种质资源为实验材料,通过对维生素E含量、含油率、皮壳率以及百粒重的测定及统计分析,试图了解向日葵种质资源中维生素E含量变异及相关数据。结果表明,在20份向日葵种质资源中,油葵的维生素E含量和含油率明显高于食葵:含油率与维生素E含量在0.01水平呈极显著正相关,含油率与百粒重在0,05水平呈显著负相关;百粒重与皮壳率在0.01水平呈极显著负相关。通过初步评价,发现3份富含天然维生素E的向日葵种质。 3) 由自由基介导的脂过氧化、酶失活或蛋白降解、胞膜破裂和遗传完整性(核酸)的损害是种子衰老的主要原因。种子在高温和高含水量情况下曝露数天诱发的加速衰老比一般衰老引起更多的生化分解;另一方面,在长期贮存条件下的低温贮存环境和种子低含水量可能使种子处于玻璃态。种子胞质的极高粘度和低分子运动能够阻止或抑制很多有害过程。尽管种子衰老的生理机制已有大量研究,但衰老过程中的主要过程和相互作用仍未完全清楚。本文报道向日葵种子在宽范围的含水量和温度条件下的脂过氧化、非酶蛋白糖基化对种子衰老的影响;同时,对种子玻璃态在长期贮存中对生化分解的阻滞作用并由此延长种子存活力也进行了探讨。
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Background: The anti-HIV-1 neutralizing antibody assay is widely used in AIDS vaccine research and other experimental and clinical studies. The vital dye staining method applied in the detection of anti-HIV-1 neutralizing antibody has been used in many laboratories. However, the unknown factor(s) in sera or plasma affected cell growth and caused protection when the tested sera or plasma was continuously maintained in cell culture. In addition, the poor solubility of neutral red in medium (such as RPMI-1640) also limited the use of this assay. Methods: In this study, human T cell line C8166 was used as host cells, and 3-(4,5-Dimethyl-2-thiazolyl)- 2,5-diphenyl-2H-tetrazolium bromide (MTT) instead of neutral red was used as vital dye. In order to avoid the effect of the unknown factor( s), the tested sera or plasma was removed by a washout procedure after initial 3 - 6 h culture in the assay. Result: This new assay eliminated the effect of the tested sera or plasma on cell growth, improved the reliability of detection of anti-HIV-1 neutralizing antibody, and showed excellent agreement with the p24 antigen method. Conclusion: The results suggest that the improved assay is relatively simple, highly duplicable, cost-effective, and well reliable for evaluating anti-HIV-1 neutralizing antibodies from sera or plasma.
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Coagulation/flocculation process was applied in the polishing treatment of molasses wastewater on a bench-scale. Important operating variables, including coagulant type and dosage, solution pH, rapid mixing conditions as well as the type and dosage of polyeletrolytes were investigated based on the maximum removal efficiencies of chemical oxygen demand (COD) and color, residual turbidity and settling characteristics of flocs. HPSEC was utilized to evaluate the removal of molecular weight fractions of melanoidins-dominated organic compounds. Experimental results indicate that ferric chloride was the most effective among the conventional coagulants, achieving 89% COD and 98% color eliminations; while aluminum sulfate was the least effective, giving COD and color reductions of 66% and 86%, respectively. In addition to metal cations, counter-ions exert significant influence on the coagulation performance since Cl--based metal salts attained better removal efficiency than SO42--based ones at the optimal coagulant dosages. Coagulation of molasses effluent is a highly pH-dependent process, with better removal efficiency achieved at lower pH levels. Rapid mixing intensity, rather than rapid mixing time, has relatively strong influence on the settling characteristics of flocs formed. Lowering mixing intensity resulted in increasing settling rate but the accumulation of floating flocs. When used as coagulant aids, synthetic polyelectrolytes showed little effects on the improvement in organic removal. On the other hand, cationic polyacrylamide was observed to substantially enhance the settleability of flocs as compared to anionic polyacrylamide. The effects of rapid mixing conditions and polymer flocculants on the coagulation performance were discussed. (C) 2009 Elsevier B.V. All rights reserved.
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The aerobic degradation of hexachlorobenzene (HCB) by an acclimated microbial community which isolated from a contaminated site and acclimated in our laboratory was investigated. The enriched microbial community was capable of biodegrading HCB when cultivated in minimal salts medium and supplied HCB as the sole carbon source. The efficiencies of microbial community in the degradation of HCB under different pH and temperatures were examined. The phylogenetic analysis for the nearly complete sequences of 16S rDNA demonstrated that the bacteria assemblage in the microbial community was dominated by Azospirillum and Alcaligenes groups.
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Hexabromocyclododecanes (HBCDs) are additive brominated flame retardants mainly used in plastics and textiles. At the present time, these compounds are found in almost all environmental and human samples. In order to evaluate the environmental safety and health risk of HBCDs, the enantiomerically pure alpha-, beta-, and gamma-HBCD were prepared using high performance liquid chromatography (HPLC) on a PM-P-CD column and the cytotoxicities of their enantiomers were evaluated in Hep G2 cells. Results from the 3-(4,5-dimethylthioazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), resazurin reduction and lactate dehydrogenase (LDH) release assays showed a good agreement that the order of cytotoxicity was gamma-HBCD >= beta-HBCD > alpha-HBCD, and that significantly lower cell viability and higher LDH release were observed in all (+)-enantiomers ((+) alpha-, (+) beta- and (+) gamma-HBCD) than the corresponding (-)-forms ((-) alpha-, (-) beta- and (-) gamma-HBCD). Additionally, the formation of reactive oxygen species (ROS) induced by these HBCD enantiomers were detected. The positive correlation between the LDH release and ROS formation demonstrated that the toxic mechanism might be mediated by oxidative damage. These results suggest that environmental and human health risks of HBCDs must be evaluated at the level of individual enantiomers. (C) 2008 Published by Elsevier Ltd.
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Polybrominated diphenyl ethers (PBDEs) are used extensively as flame-retardants and are ubiquitous in the environment and in wildlife and human tissue. Recent studies have shown that PBDEs induce neurotoxic effects in vivo and apoptosis in vitro. However, the signaling mechanisms responsible for these events are still unclear. In this study, we investigated the action of a commercial mixture of PBDEs (pentabrominated diphenyl ether, DE-71) on a human neuroblastoma cell line, SK-N-SH. A cell viability test showed a dose-dependent increase in lactate dehydrogenase leakage and 3-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyl-tetrazolium bromide reduction. Cell apoptosis was observed through morphological examination, and DNA degradation in the cell cycle and cell apoptosis were demonstrated using flow cytometry and DNA laddering. The formation of reactive oxygen species was not observed, but DE-71 was found to significantly induce caspase-3, -8, and -9 activity, which suggests that apoptosis is not induced by oxidative stress but via a caspase-dependent pathway. We further investigated the intracellular calcium ([Ca2+](i)) levels using flow cytometry and observed an increase in the intracellular Ca2+ concentration with a time-dependent trend. We also found that the N-methyl d-aspartate (NMDA) receptor antagonist MK801 (3 mu M) significantly reduced DE-71-induced cell apoptosis. The results of a Western blotting test demonstrated that DE-71 treatment increases the level of Bax translocation to the mitochondria in a dose-dependent fashion and stimulates the release of cytochrome c (Cyt c) from the mitochondria into the cytoplasm. Overall, our results indicate that DE-71 induces the apoptosis of ([Ca2+](i)) in SK-N-SH cells via Bax insertion, Cyt c release in the mitochondria, and the caspase activation pathway.
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A simple, sensitive, and accurate method for determination of polybrominated diphenyl ethers (PBDEs) in soil has been developed based on headspace solid-phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry (GC-MS). Permethylated-beta-cyclodextrin/hydroxyl-termination silicone oil (PM-beta-CD/OH-TSO) fiber was first prepared by sol-gel technology and employed in SPME procedure. By exploiting the superiorities of sot-gel coating technique and the advantages of the high hydrophobic doughnut-shaped cavity of PM-beta-CD, the novel fiber showed desirable operational stability and extraction ability. After optimization on extraction conditions like water addition, extraction temperature, extraction time, salts effect, and solvents addition, the method was validated in soil samples, achieving good linearity (r>0.999), precision (R.S.D. < 10%), accuracy (recovery>78%), and detection limits (S/N =3) raging from 13.0 to 78.3 pg/g. (c) 2007 Published by Elsevier B.V.
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Several assay methods were screened for viability assessment in cyanobacteria using Microcystis aeruginosa FACHB 905. Compared with fluorescent diacetate (FDA), Evan's Blue and autofluorescence, the 3-[4,5-dimethylthiazol-2-yl]2,5-diphenyl tetrazolium bromide (MTT) assay, which was based on the ability of viable cells to reduce MTT to formazan, was found to be reliable and was selected for further study. MTT concentration, incubation time and temperature were optimized for M. aeruginosa. Improvements to the sensitivity and reproducibility of the MTT assay included performing it in the dark to reduce the effects of formazan light sensitivity when extracted in DMSO. Another improvement involved collecting viability data by cell by counting rather than colourimetrically, which was concluded from the fact that oxidoreductase activity, responsible for MTT reduction, would elevate or decrease under stress conditions. Half-life of oxidoreductase in dead cell was calculated to be 3 h. The MTT assay was also found to be applicable to other cyanobacteria and diatoms, including field samples, but not for algae belonging to Chlorophyta, Euglenophyta, Pyrrophyta or Chrysophyta. Based on the above results, we proposed an optimized procedure for the MTT method on Microcystis strains. The use of this assay may be of importance to better understand the dynamics of bloom and the fate of Microcystis under natural or disturbed conditions.
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Polybrominated diphenyl ethers (PBDEs) are an important class of halogenated organic brominated flame retardants. Because of their presence in abiotic and biotic environments widely and their structural similarity to polychlorinated biphenyls (PCBs), concern has been raised on their possible adverse health effects to humans. This study was designed to determine the anti-proliferative, apoptotic properties of decabrominated diphenyl ether (PBDE-209), using a human hepatoma Hep G2 line as a model system. Hep G2 cells were cultured in the presence of PBDE-209 at various concentrations (1.0-100.0 mu mol/L) for 72 h and the percentage of cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The results showed that PBDE-209 inhibited the cells viability in time and concentration-dependent characteristics at concentrations (10.0-100.0 mu mol/L). We found that anti-proliferative effect of PBDE-209 was associated with apoptosis on Hep G2 cells by determinations of morphological changes, cell cycle and apoptosis. Mechanism study showed that PBDE-209 could increase the generation of intracellular reactive oxygen species (ROS) concentration-dependently. Antioxidant N-acetylcyteine partially inhibited the increase of ROS. The mechanism for its hepatoma-inhibitory effects was the induction of cellular apoptosis through ROS generation. In addition, activity of lactate dehydrogenase (LDH) release increased when the cells incubated with PBDE-209 at various concentrations and times. These results suggested that PBDE-209 had the toxicity activity of anti-proliferation and induction of apoptosis in tumor cells in vitro. (c) 2007 Elsevier Ireland Ltd. All rights reserved.
Resumo:
Gamete release is an essential event in artificial seeding of the economic brown seaweed, Hizikia fusiforme. Mass egg release occurred in the dark, with few eggs being discharged in the light. Release of eggs was elicited with eight practical salinity units (one PSU = 1 g sea salts l(-1)) and was inhibited by salinity levels > 32 PSU. Egg release was optimal at 23 degrees C, and was decreased by 72% in agitated seawater compared to unstirred seawater. Inhibitors of photosynthesis and ions channels suppressed egg release, indicating that this process was physiologically associated with photosynthetic activity and ion transport.